CAJ | 학술논문

目的探讨热休克蛋白47( HSP47)在肾小管间质纤维化中的作用及其可能机制.方法常规培养人肾小管上皮细胞( HK-2),分为对照组、转化生长因子β1( TGF-β1)组、HSP47 - siRNA组.RT-PCR检测HSP47、胶原Ⅳ、纤连蛋白(FN)、组织型纤溶酶原激活物抑制剂1 (PAI-1)的mRNA表达.Western印迹检测HSP47、胶原Ⅳ、FN蛋白表达.ELISA检测PAI-1的蛋白量.结果 HK-2细胞有HSP47表达.不同浓度TGF-β1(0、2.5、5、10 μg/L)干预不同时间(12、24、48 h)时,HSP47基因和蛋白表达呈浓度和时间依赖性增高,10 μg/L TGF-β1干预HK-2细胞48 h时,HSP47 mRNA和蛋白表达最强.不同浓度TGF-β1(0、5、10 μg/L)干预HK-2不同时间(12、24、48 h)时,胶原Ⅳ、FN、PAI-1 mRNA和蛋白表达亦呈浓度和时间依赖性增高,10 μg/L TGF-β1作用48 h时,3者mRNA和蛋白表达最强.与TGF-β1组比较,HSP47-siRNA组的HSP47、胶原Ⅳ、FN、PAI-1 mRNA和蛋白表达都明显下调.结论 HSP47可促进肾小管间质纤维化,其机制可能与上调胶原Ⅳ、FN、PAI-1表达有关.
목적 탐토열휴극단백47( HSP47)재신소관간질섬유화중적작용급기가능궤제.방법 상규배양인신소관상피세포( HK-2),분위대조조、전화생장인자β1( TGF-β1)조、HSP47 - siRNA조.RT-PCR검측HSP47、효원Ⅳ、섬련단백(FN)、조직형섬용매원격활물억제제1 (PAI-1)적mRNA표체.Western인적검측HSP47、효원Ⅳ、FN단백표체.ELISA검측PAI-1적단백량.결과 HK-2세포유HSP47표체.불동농도TGF-β1(0、2.5、5、10 μg/L)간예불동시간(12、24、48 h)시,HSP47기인화단백표체정농도화시간의뢰성증고,10 μg/L TGF-β1간예HK-2세포48 h시,HSP47 mRNA화단백표체최강.불동농도TGF-β1(0、5、10 μg/L)간예HK-2불동시간(12、24、48 h)시,효원Ⅳ、FN、PAI-1 mRNA화단백표체역정농도화시간의뢰성증고,10 μg/L TGF-β1작용48 h시,3자mRNA화단백표체최강.여TGF-β1조비교,HSP47-siRNA조적HSP47、효원Ⅳ、FN、PAI-1 mRNA화단백표체도명현하조.결론 HSP47가촉진신소관간질섬유화,기궤제가능여상조효원Ⅳ、FN、PAI-1표체유관.
Objective To study the role of hot shock protein (HSP) 47 in tubulointerstitial fibrosis induced by transforming growth factor β1 (TGF-β1),and to explore its possible mechanism. Methods Human proximal tubular epithelial cells (HK-2) were divided into three groups: control,TGF-β1 and HSP47siRNA. The expressions of HSP47, collagen Ⅳ,fibronectin (FN),plasminogen activator inhibitor 1 (PAI-1) mRNA and HSP47,collagen Ⅳ,FN protein were detected by RT-PCR and Western blotting respectively.PAl-1 protein was detected by ELISA. Results HK-2 expressed HSP47 in normal medium. The mRNA and protein expressions of HSP47 up-regulated in concentration- and time-dependent manner in HK-2 cells induced with increasing concentrations of TGF-β1 (0,2.5,5,10 μg/L) and with prolong times (12,24,48 h),and peaked at 10 μg/L TGF-β1 for 48 h.Similar phenomena was observed in the mRNA and protein expressions of collagen Ⅳ, FN, PAI-1 in HK-2 cells induced by increasing concentrations of TGF-β1 (0,2.5,5,10 μg/L) at different time points (12,24,48 h),and peaked at 10 μg/L TGF-β1 for 48 h.HSP47 siRNA could significantly reduce the up-regulation of mRNA and protein expressions of HSP47,collagen Ⅳ,FN,PAI-1 in HK-2 cells induced by TGF-β1.Conclusion HSP47 can promote renal tubulointerstitial fibrosis maybe through the regulation of the expressions of collagen Ⅳ,FN,PAI-1.